Em18.3重组蛋白的表达及其免疫诊断特性的研究  

Expression and serological evaluation of diagnostic antigen Em18 for alveolar echinococcosis

作　　者：林仁勇[1] 张春桃[1] 温浩[1] 王俊芳[1] 王星[1] 王俨[1] 卢晓梅[1] 张静萍[1] 张琰[1] 无 

机构地区：[1]新疆医科大学第一附属医院,新疆包虫病基础医学重点实验室,新疆乌鲁木齐830054 [2]新疆医科大学基础医学院免疫学教研室

出　　处：《新疆医科大学学报》2007年第4期332-335,共4页Journal of Xinjiang Medical University

基　　金：国家自然科学基金(NO.30360097);新疆维吾尔自治区高校科研计划创新研究群体基金(XJEDU2004G10);新疆重点实验室开放课题基金(XJDX0202-2003-01)资助项目

摘　　要：目的:在大肠杆菌中表达和纯化截短的泡球蚴rEm18.3抗原重组蛋白,对其免疫诊断特性进行鉴定。方法:IPTG诱导pET41a-Em18.3原核表达质粒,表达和纯化rEm18.3-GST重组蛋白,SDS-PAGE电泳及West-ern blot法对其进行初步鉴定,ELISA法确定其免疫诊断特性。结果:rEm18.3-GST重组蛋白得到成功表达,SDS-PAGE电泳显示相对分子量为45kDa。Western blot结果表明,rEm18.3-GST重组蛋白能被AE病人阳性血清识别。对56例多房棘球蚴病(AE)、88例细粒棘球蚴病(CE)、24例其他寄生虫病、24例其他疾病和24例健康者共236份血清进行ELISA检测结果显示,对AE血清诊断的敏感性为10.7%,特异性为99.4%,阳性预测值85.7%,阴性预测值78.1%。结论:rEm18.3-GST免疫诊断价值较低,Em18抗原重要的表位可能位于1-40位氨基酸。Objective： To express and serologically evaluate the rEm18-3 antigen gene of Echinococcus multilocularis for diagnostic purpose. Methods： The constructed plasmid pET41a-Em18.3 was transferred into E. coli BL21 （DE3） for expression. The recombinant proteins were purified with His-Binding agarose by affinity chromatography. Twohundred and thirty six sera were used for evaluating diagnostic value of the recombinant Em18.3. Resulits： The expressed rEm18. 3-GST recombinant protein ould be detected as a band of 45 kDa by SDS-PAGE and Western blot result confirmed that the recombinant protein could specifically react with the serum samples from patients with alveolar echinococcosis （AE）. The molecular weight of the expression protein was 45 kDa. Serological evaluation by ELISA was carried out using sera from 56 patients with alveolar echinococcosis（AE）, 88 with cystic echinococcosis（CE）, 24 from other parasite patients, 44 from non-parasite patients, and 24 from healthy persons （HP）. The results showed an overall sensitivity of 10.7 % and specificity of 99.4%, positive predictive value 85.7 % and negative predictive value 78.1%. Conclusion： The diagnostic value of rEm18.3 was not high.

关 键 词：泡球蚴 Em18重组蛋白 免疫检测 

分 类 号：R532.32[医药卫生—内科学] R392.7[医药卫生—临床医学]